Monitoring Drug Target Engagement in Cells and Tissues Using the Cellular Thermal Shift Assay
Abstract
Drug Targeting: Drug efficacy depends on the extent of binding to a cellular target (often a protein) with adverse effects caused by excessive target binding or by off-target binding. Engagement of a target protein inside cells is influenced by the effective drug concentration and by factors that regulate the protein conformation, making it difficult to predict efficacy based on in vitro affinity studies. Martinez Molina et al. (p. 84 ) took advantage of the shift in protein thermal stability caused by drug binding to directly monitor target protein-drug interactions in cells. The method was used to monitor drug target engagement in cancer cells and in mouse livers and kidneys.
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Both CETSA and BiTSA are effective methods for assessing the thermal stability of ApoE isoforms and variants.
“In this study, we demonstrate that both CETSA 42 and the BiTSA 45 assay are effective methods for assessing the thermal stability of ApoE isoforms and variants.”
From Utilizing a cell culture based novel cellular thermal shift assay to understand the isoform-dependent thermal stability of ApoE variants (2026) · cites this paper as [42]
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“Next, we evaluated this model through a cell-based target engagement by SH6 using the cellular thermal shift assay (CETSA) (Jafari et al, 2014; Martinez Molina et al, 2013).”
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- Authors
- Molina, D. M., Jafari, R., Ignatushchenko, M., Seki, T., Larsson, E. A., Dan, C., Sreekumar, L., Cao, Y., & Nordlund, P.
- Affiliations
- Karolinska Institutet (SE)
- Nanyang Technological University (SG)
- School of Biological Sciences, Nanyang Technological University, 61 Nanyang Drive, Singapore 639798, Singapore.
- Linköping University (SE)
- Department of Medical Biochemistry and Biophysics, Karolinska Institute, Scheeles väg 2, SE-17177 Stockholm, Sweden.
- Venue
- Science
- Published
- 2013-07-05
- Volume
- 341
- Issue
- 6141
- Pages
- 84-87
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W1993251792
- Cited by
- 2,261
- References
- 3
- Citations by year
- 2026: 299 · 2025: 268 · 2024: 299 · 2023: 242 · 2022: 204 · 2021: 203 · 2020: 194 · 2019: 137 · 2018: 126 · 2017: 106 · 2016: 88 · 2015: 53 · 2014: 33 · 2013: 6
- FWCI
- 38.90
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Science (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Chemistry › Physical and Theoretical Chemistry › thermodynamics and calorimetric analyses
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Interaction Studies and Fluorescence Analysis
- Keywords
- cellular thermal shift assay, drug-target engagement, protein thermal stability, off-target effects, drug resistance, drug transport, tissue drug distribution, cancer cell lines, drug target validation
- MeSH terms
- Antimetabolites, Antineoplastic, Dose-Response Relationship, Drug, Pharmaceutical Preparations, Folic Acid Antagonists, Hot Temperature, Humans, Kidney, Ligands, Liver, Protein Binding, Proteins, Quinazolines, Thiophenes, Tissue Distribution, Drug Monitoring, Cell Line, Tumor, Protein Stability, Molecular Targeted Therapy
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